A Review of Fluorescent Sensor Development for Ultrasensitive Per- and Polyfluoroalkyl Compound Analysis in Water: Toward in Situ Monitoring Platforms Concetta Esposito, Angela Maria Cusano, Tania Mariastella Caputo, Anna Aliberti, Andrea Cusano ACS Materials Letters, 2026 Anthropogenic per- and polyfluoroalkyl substances (PFAS), valued for their chemical stability, are widely used in industrial and consumer products. Their persistence leads to bioaccumulation, particularly in water, posing significant risks to human health via contaminated water, food, and PFAS-treated products. Although regulations exist, gold standard analysis is time-consuming, complex, and costly, limiting real-time and in situ monitoring. Efficient, field-deployable detection technologies are urgently needed. This review reports an up-to-date and critical assessment of fluorescence-based sensors, often harnessing engineered nanomaterials, for the detection of PFAS in water with a view to portable systems for in situ monitoring. We discuss the photophysical and chemical principles of these sensors and evaluate key performance metrics─including sensitivity, selectivity, response time, and sample preparation─that affect operational efficiency and field portability. Despite challenges such as matrix interference and sensitivity limits, emerging nanomaterial designs and sensor architectures offer promise for robust, practical continuous in situ PFAS monitoring.
Engineered anti-HER2 drug delivery nanosystems for the treatment of breast cancer Silvia Vanni, Tania Mariastella Caputo, Angela Maria Cusano, Alessandro De Vita, Andrea Cusano, Claudia Cocchi, Chiara Mulè, Sofia Principe, Chiara Liverani, Giorgia Celetti, Alberto Micco, Chiara Spadazzi, Giacomo Miserocchi, Toni Ibrahim, Laura Mercatali, Anna Aliberti Nanoscale, 2025 Schematic representation of clinical and pre-clinical workflow settings for light controlled release of PLGA NPs for breast cancer treatment. Created in BioRender.com. Vanni, S. (2025) https://BioRender.com/r11b297.
PLGA nanoparticles for capsaicin delivery: enhanced encapsulation efficiency and pro-apoptotic activity in HEPG2 cells Chiara Mulè, Tania Mariastella Caputo, Antonio Montefusco, Antonio Massimiliano Romanelli, Ivana Caputo, Gaetana Paolella, Anna Aliberti, Andrea Cusano Frontiers in Bioengineering and Biotechnology, 2025 IntroductionCapsaicin (trans-8-methyl-N-vanillyl-6-nonenamide) (Cap) is a lipophilic alkaloid derived from Capsicum annuum. It was observed that Cap has an antitumoral activity in several cancer types, in particular in liver, colon and breast cancer. Actually, the use of Cap in the cancer therapy is limited by its very low bioavailability, a short half-life and side effects as mouth and stomach irritations and burning sensation. To overcome these limitations, the Cap has been encapsulated in carriers in order to reduce the adverse effect and to help the delivery in the cancer cells. In this study, we synthesized Poly(lactic co-glycolic acid) (PLGA) nanoparticles (NPs) to encapsulate Cap (PLGA-Cap), optimizing the synthetic strategy and improving its efficiency and safety. This is the first time that PLGA-Cap NPs was tested on HepG2 cells line for Hepatocellular carcinoma (HCC) therapy.MethodsNPs are characterized by Dynamic Light Scattering (DLS), Fourier transform infrared spectroscopy (FTIR), Morphological analysis by scanning transmission electron microscopy (STEM) and Reverse-Phase High Liquid Chromatography (RP-HPLC) to study their physicochemical properties and the best condition in terms of size, PDI and encapsulation efficiency. In vitro biological MTT assay was performed on HepG2 cells to observe the cell proliferation in response to PLGA-Cap. The apoptosis induced by Cap was evaluated the enzymatic activity of caspase 3, Bcl2 and Bax expression by Western blot and ROS activity.Results and DiscussionOur preparation showed the highest Encapsulation Efficiency (96%) reported by the literature, showing an improvement of 21% compared to what is actually reported. In vitro experiments revealed that PLGA-Cap formulation induced similar biological effects in terms of cell viability compared to free Cap. Moreover, HepG2 cancer cells treated with PLGA-Cap exhibited increased caspase 3 activity respect to those treated with free Cap.ConclusionIn conclusion we demonstrated that our preparation showed an improvement in encapsulation parameters and in pro-apoptotic and anticancer activity in HepG2 cells.
Fluorescein Isothiocyanate Labelled PCL-PEG-PCL Copolymer as Delivery System of Capsaicin Nancy Ferrentino, Tania Mariastella Caputo, Angela Maria Cusano, Anna Aliberti, Andrea Cusano, Daniela Pappalardo Chemnanomat, 2024 Amphiphilic block copolymers, made of biocompatible polycaprolactone (PCL) and polyethylene glycol (PEG), due to their ability to self‐assemble in water into nanoscopic micelles, have been largely exploited for drug delivery systems (DDS). This study introduces a novel approach by synthesizing a fluorescein isothiocyanate FITC‐labelled PCL‐PEG‐PCL triblock copolymer, with the aim to develop a drug delivery system for natural bioactive. As a proof of concept, the FITC‐labelled PCL‐PEG‐PCL copolymer is applied for the preparation of micelles encapsulating into the core capsaicin (CP), a pungent alkaloid found in chili peppers with diverse therapeutic applications. Challenges associated with CP′s solubility, bioavailability, and stability are addressed using this DDS. Comprehensive characterization of FITC‐labelled copolymer is conducted using a range of analytical techniques, including nuclear magnetic resonance (NMR), dynamic light scattering (DLS), high‐performance liquid chromatography (HPLC), Fourier‐transform infrared spectroscopy (FTIR), fluorescence, and confocal laser scanning microscopy (CLSM). Key properties such as critical micelle concentration, CP loading, and release behavior are thoroughly investigated and compared with the characteristics of the unlabeled parent copolymer. This research pioneers the investigation of PCL‐PEG‐PCL triblock copolymers for CP delivery, along with the use of FITC‐labelled variants, opening new avenues for research in drug delivery and nanomedicine.
Precision Point-Of-Care in Drug Delivery: Empowering Innovations with Optical Fiber assisted by Microfluidics Tania Mariastella Caputo, Anna Aliberti, Angela Maria Cusano, Chiara Mulè, Alberto Micco, Andrea Cusano Proceedings of SPIE the International Society for Optical Engineering, 2024 Recent advancements in loco-regional therapy are revolutionizing the point-of-care (POC) drug delivery field, enhancing the convenience, comfort, and effectiveness of these devices for patients. New anticancer drugs have improved the therapy, but there is still significant work to be done in order to selectively target tumour cells. To bridge this gap, here, we present a minimally invasive tool based on optical fibers (OFs) integrated in a microfluidic device for light-triggered loco regional delivery of drug-loaded particles. Specifically, we, here, selected the Lab on Fiber (LOF) technology as an attractive option to guide light and trigger the drug release. LOF, indeed, enables precise drug dosing and targeted transport, laying the foundation for mini-invasive platforms in light-activated loco-regional drug delivery. The platform includes drug-loaded carriers covalently attached on the OF surface through a photocleavable linker. The OF platform has been designed to spread light from the core region towards the cladding area in order to achieve enough power density to photo-cleave the linker and release the carrier. To this aim, side-emitting and core-offset OFs optrodes have been designed, fabricated and characterized achieving power density and scattering efficiency of 30% and 5mW/cm<sup>2</sup> and 24.9% and 4.64mW/cm<sup>2</sup>. OFs were integrated into a microfluidic device and the particle release upon light activation was quantified. The side-emitting OF microfluidic system, designed for Hepatocellular cancer therapy, released 2.60μg/ml of carriers, while the core-offset OF microfluidic system, for Breast cancer treatment, released 1.06μg/ml of carriers. This approach holds potential for improving cancer treatment outcomes
Development of High-Loading Trastuzumab PLGA Nanoparticles: A Powerful Tool Against HER2 Positive Breast Cancer Cells Tania Mariastella Caputo, Giovannina Barisciano, Chiara Mulè, Angela Maria Cusano, Anna Aliberti, Livio Muccillo, Vittorio Colantuoni, Lina Sabatino, Andrea Cusano International Journal of Nanomedicine, 2023 Background Trastuzumab, a therapeutic monoclonal antibody directed against HER2, is routinely used to treat HER2-positive breast cancer with a good response rate. However, concerns have arisen in the clinical practice due to adverse side effects. One way to overcome these limitations is to encapsulate trastuzumab in nanoparticles to improve cytotoxic activity, increase intracellular drug concentrations, escape the immune system and avoid systemic degradation of the drug in vivo. Methods A double emulsion method was used to encapsulate trastuzumab into poly(lactic-co-glycolic) nanoparticles, effective for their biocompatibility and biodegradability. These nanocarriers, hereafter referred to as TZPs, were characterised in terms of size, homogeneity, zeta potential and tested for their stability and drug release kinetics. Finally, the TZPs cytotoxicity was assessed in vitro on the HER2 positive SKBR3 breast cancer cell line and compared to free trastuzumab. Results The TZPs were stable, homogeneous in size, with a reduced zeta potential. They showed higher encapsulation efficiency and drug loading, a prolonged trastuzumab release kinetics that retained its physicochemical properties and functionality. TZPs showed a stronger cytotoxicity and increased apoptosis than similar doses of free trastuzumab in the cell line analysed. Confocal microscopy and flow cytometry assessed TZPs and trastuzumab cellular uptake while Western blot evaluated downstream signalling, overall HER2 content and shedding. Conclusion TZPs exert more robust effects than free trastuzumab via a dual mode of action: TZPs are taken up by cells through an endocytosis mechanism and release the drug intracellularly for longer time. Additionally, the TZPs that remain in the extracellular space release trastuzumab which binds to the cognate receptor and impairs downstream signalling. This is the sole modality used by free trastuzumab. Remarkably, half dose of TZPs is as efficacious as the highest dose of free drug supporting their possible use for drug delivery in vivo.
Sorafenib-Loaded PLGA Carriers for Enhanced Drug Delivery and Cellular Uptake in Liver Cancer Cells Tania Mariastella Caputo, Angela Maria Cusano, Sofia Principe, Paola Cicatiello, Giorgia Celetti, Anna Aliberti, Alberto Micco, Menotti Ruvo, Maria Tagliamonte, Concetta Ragone, Michele Minopoli, Maria Vincenza Carriero, Luigi Buonaguro, Andrea Cusano International Journal of Nanomedicine, 2023 Introduction Currently, conventional treatments of hepatocellular carcinoma (HCC) are not selective enough for tumor tissue and lead to multidrug resistance and drug toxicity. Although sorafenib (SOR) is the standard first-line systemic therapy approved for the clinical treatment of HCC, its poor aqueous solubility and rapid clearance result in low absorption efficiency and severely limit its use for local treatment. Methods Herein, we present the synthesis of biodegradable polymeric Poly (D, L-Lactide-co-glycolide) (PLGA) particles loaded with SOR (PS) by emulsion-solvent evaporation process. The particles are carefully characterized focusing on particle size, surface charge, morphology, drug loading content, encapsulation efficiency, in vitro stability, drug release behaviour and tested on HepG2 cells. Additionally, PLGA particles have been coupled on side emitting optical fibers (seOF) integrated in a microfluidic device for light-triggered local release. Results PS have a size of 248 nm, tunable surface charge and a uniform and spherical shape without aggregation. PS shows encapsulation efficiency of 89.7% and the highest drug loading (8.9%) between the SOR-loaded PLGA formulations. Treating HepG2 cells with PS containing SOR at 7.5 µM their viability is dampened to 40%, 30% and 17% after 48, 129 and 168 hours of incubation, respectively. Conclusion The high PS stability, their sustained release profile and the rapid cellular uptake corroborate the enhanced cytotoxicity effect on HepG2. With the prospect of developing biomedical tools to control the spatial and temporal release of drugs, we successfully demonstrated the potentiality of seOF for light-triggered local release of the carriers. Our prototypical system paves the way to new devices integrating microfluidics, optical fibers, and advanced carriers capable to deliver minimally invasive locoregional cancer treatments.
Human Serum Albumin Nanoparticles as a Carrier for On-Demand Sorafenib Delivery Tania Mariastella Caputo, Angela Maria Cusano, Menotti Ruvo, Anna Aliberti, Andrea Cusano Current Pharmaceutical Biotechnology, 2022 Background: Drug delivery systems based on Human Serum Albumin (HSA) have been widely investigated due to their capability to interact with several molecules together with their nontoxicity, non-immunogenicity and biocompatibility. Sorafenib (SOR) is a kinase inhibitor used as the firstline treatment in hepatic cancer. However, because of its several intrinsic drawbacks (low solubility and bioavailability), there is a growing need for discovering new carriers able to overcome the current limitations. Objective: To study HSA particles loaded with SOR as a thermal responsive drug delivery system. Method: A detailed spectroscopy analysis of the HSA and SOR interaction in solution was carried out in order to characterize the temperature dependence of the complex. Based on this study, the synthesis of HSA particles loaded with SOR was optimized. Particles were characterized by Dynamic Light Scattering, Atomic Force Microscopy and by spectrofluorometer. Encapsulation efficiency and in vitro drug release were quantified by RP-HPLC. Results: HSA particles were monodispersed in size (≈ 200 nm); encapsulation efficiency ranged from 25% to 58%. Drug release studies that were performed at 37 °C and 50 °C showed that HS5 particles achieved a drug release of 0.430 μM in 72 hours at 50 °C in PBS buffer, accomplishing a 4.6-fold overall SOR release enhancement following a temperature increase from 37 °C to 50 °C. Conclusion: The system herein presented has the potential to exert a therapeutic action (in the nM range) triggering a sustained temperature-controllable release of relevant drugs.
Hydrogel microparticles for fluorescence detection of mirna in mix-read bioassay Alessia Mazzarotta, Tania Mariastella Caputo, Edmondo Battista, Paolo Antonio Netti, Filippo Causa Sensors, 2021 Herein we describe the development of a mix-read bioassay based on a three-dimensional (3D) poly ethylene glycol—(PEG)-hydrogel microparticles for the detection of oligonucleotides in complex media. The key steps of hydrogels synthesis and molecular recognition in a 3D polymer network are elucidated. The design of the DNA probes and their density in polymer network were opportunely optimized. Furthermore, the diffusion into the polymer was tuned adjusting the polymer concentration and consequently the characteristic mesh size. Upon parameters optimization, 3D-PEG-hydrogels were synthetized in a microfluidic system and provided with fluorescent probe. Target detection occurred by double strand displacement assay associated to fluorescence depletion within the hydrogel microparticle. Proposed 3D-PEG-hydrogel microparticles were designed for miR-143-3p detection. Results showed 3D-hydrogel microparticles with working range comprise between 10−6–10−12 M, had limit of detection of 30 pM and good specificity. Moreover, due to the anti-fouling properties of PEG-hydrogel, the target detection occurred in human serum with performance comparable to that in buffer. Due to the approach versatility, such design could be easily adapted to other short oligonucleotides detection.
Small oligonucleotides detection in three-dimensional polymer network of dna-peg hydrogels Alessia Mazzarotta, Tania Mariastella Caputo, Luca Raiola, Edmondo Battista, Paolo Antonio Netti, Filippo Causa Gels, 2021 The control of the three-dimensional (3D) polymer network structure is important for permselective materials when specific biomolecule detection is needed. Here we investigate conditions to obtain a tailored hydrogel network that combines both molecular filtering and molecular capture capabilities for biosensing applications. Along this line, short oligonucleotide detection in a displacement assay is set within PEGDA hydrogels synthetized by UV radical photopolymerization. To provide insights on the molecular filter capability, diffusion studies of several probes (sulforhodamine G and dextrans) with different hydrodynamic radii were carried out using NMR technique. Moreover, fluorometric analyses of hybridization of DNA oligonucleotides inside PEGDA hydrogels shed light on the mechanisms of recognition in 3D, highlighting that mesh size and crowding effect greatly impact the hybridization mechanism on a polymer network. Finally, we found the best probe density and diffusion transport conditions to allow the specific oligonucleotide capture and detection inside PEGDA hydrogels for oligonucleotide detection and the filtering out of higher molecular weight molecules.
